Clinical subtypes and molecular basis of epidermolysis bullosa in Kuwait.

BACKGROUND: Epidermolysis bullosa (EB) is a clinically and genetically heterogeneous blistering skin disease, but in countries such as Kuwait, there are very limited data on the clinical and molecular pathology of EB. To improve understanding of EB in Kuwait, we report the experience of a local tertiary referral center over a 17.5 year period (January 2000-June 2017) in establishing clinical and molecular diagnoses. METHODS: Review of hospital records and diagnostic reports. Individual cases were diagnosed by combinations of clinical assessment, skin biopsy (immunohistochemistry and transmission electron microscopy), Sanger sequencing of EB genes, and whole exome sequencing. RESULTS: Fifty-four families with EB were registered with the clinic over this period, 41 of whom (84 patients) participated in diagnostic studies. Thirty-seven of these 41 families had consanguineous marriages; 34 had recessive forms of EB, while only seven had dominant subtypes. Recurrent mutations were observed in epidermal dystonin, transglutaminase 5, and type VII collagen. CONCLUSIONS: The prevalence of EB in Kuwait is approximately three times that of internationally cited rates with an over-representation of autosomal recessive variants. Establishing the molecular basis of EB in Kuwait with accurate diagnostic subtyping provides a basis for determining healthcare requirements and improving patient management of EB.

Hypercoagulable state and methylenetetrahydrofolate reductase (MTHFR) C677T mutation in patients with beta-thalassemia major in Kuwait.

INTRODUCTION: Patients with thalassemia major often present with a hypercoagulable state, the pathogenesis of which is still not understood. MATERIALS AND METHODS: This study evaluates the risk factors for hypercoagulability in 50 beta-thalassemia major patients and 50 healthy controls. Fasting total homocysteine, protein C (PC), protein S (PS), antithrombin (AT), activated protein C resistance (APCR) and lupus anticoagulant (LA) were assessed. MTHFR C677T mutation was determined. RESULTS: Significant reductions in PC, PS and AT were noted in patients. Only 4% of the patients had hyperhomocysteinemia. Thirty-two percent of the patients were heterozygous and 4% were homozygous for MTHFR C677T mutation. CONCLUSION: The natural coagulation inhibitors PC, PS and AT were significantly reduced in patients with beta-thalassemia major and were thus important risk factors for the hypercoagulable state, but hyperhomocysteinemia and MTHFR mutation do not seem to be significant risk factors for thromboembolic events.

Screening and validation of dinucleotide repeats in intron 1 of the human EGFR gene and its paralog in the HER2 gene.

Overexpression or dysfunction of EGFR and other ErbBs is known to be involved in many human cancers. The first intron of several genes, including ErbBs, has an important regulatory function in transcription. In intron 1 of the EGFR gene, simple CA repeats (CA-SSR) have been found to be associated with the level of transcriptional modulation of the EGFR both in vitro and in vivo. The aim of this work was to screen for conserved dinucleotide repeats located in introns of the human ErbB genes. Pairwise BLAST was used to identify paralogs to intron 1 of EGFR in the HER2 gene. Dinucleotide tandem repeats were searched in intron sequences using the Tandem Repeat Software to restrict detection of dinucleotide microsatellites containing at least 10 repeats. With multiple alignment, short conserved DNA sequences should also be detected, revealing the presence of potential regulatory elements near CA repeats. We found that the nearest homolog to intron 1 in the EGFR gene is intron 4 of the HER2 gene. The experimental validation of four predicted short tandem repeats (STRs) (three dinucleotide repeats in intron 1 of the EGFR and one in intron 4 of the HER2 gene) by genotyping of about 100 controls showed that these STRs are polymorphic. In a case-control study to test the association of the four new polymorphic STRs with breast cancer, a significant allelic association was found for all STRs (P < 0.001). These results suggest that the role of transcriptional regulation of CA repeats in intron 1 of the EGFR gene might be conserved in other ErbB genes.

Lymphocyte subsets in healthy adult Kuwaiti Arabs with relative benign ethnic neutropenia.

Relative and absolute neutropenia is frequently seen in the healthy adult Kuwaiti Arab population. Fluorescent monoclonal antibody labelling followed by flow cytometry was used to determine the lymphocyte subsets in 48 normal healthy individuals in the Kuwaiti adult population (24 males and 24 females, age 17-59 years) with relative or absolute neutropenia, and this was compared to age-matched controls (64 males and 63 females). The mean haemoglobin levels were 13.6+/-1.5 and 13.7+/-1.5 g/dl in the neutropenic and control groups, respectively. White blood cell counts, absolute neutrophil and lymphocyte counts in neutropenic individuals with the corresponding reference range, taken from the control subjects (in parenthesis) were: WBC, 6.7+/-1.6 x 10(9)/l (4-10.4 x 10(9)/l), neutrophils, 2.7+/-0.8 x 10(9)/l (1.87-6.63 x 10(9)/l), lymphocytes, 3.3+/-0.9 x 10(9)/l (1.4-3.62 x 10(9)/l). Absolute values of lymphocytes, CD2+, CD3+, CD19+, CD4+, CD8+, HLADR+ and CD45RA+ cells were significantly higher in the neutropenic group. The range of values with the corresponding reference ranges, in parenthesis, were: CD2+, 1.61-4.30 x 10(9)/l (0.95-2.99 x 10(9)/l), CD3+, 1.37-4.16 x 10(9)/l (0.83-2.71 x 10(9)/l), CD19+, 0.16-1.09 x 10(9)/l (0.05-0.61 x 10(9)/l), CD4+, 0.70-2.89 x 10(9)/l (0.45-1.65 x 10(9)/l), CD8+, 0.57-1.80 x 10(9)/l (0.29-1.17 x 10(9)/l), HLADR+ 0.27-1.74 x 10(9)/l (0.02-0.62 x 10(9)/l), CD45RA, 0.90-4.63 x 10(9)/l (0.34-2.05 x 10(9)/l), respectively. The levels of natural killer cells, CD56+ cells were significantly lower compared to controls while the values of memory T lymphocytes, CD45RO+ were comparable to controls. These results indicate that difference in the leukocyte subpopulations may also be indicative of differences in the lymphocyte subpopulations and that reference ranges for these cell types in healthy neutropenic and non-neutropenic individuals should be established.

Reference values of lymphocyte subsets in the normal healthy adult Kuwaiti Arab population.

Fluorescent monoclonal antibody labelling followed by a lysed whole blood method and flow cytometry was used to determine the lymphocyte subpopulations in 127 (64 males and 63 females) normal healthy individuals in the adult (age 18-59 years) Kuwaiti population. Relative percentages and absolute values of CD2+, CD3+, CD19+, CD4+, CD8+, HLADR+, CD56+, CD45RO+, and CD45RA+ cells were determined. The reference ranges were CD2+, 73-92% (0.95-2.99 x 10(9) per l), CD3+, 64-85% (0.83-2.71 x 10(9) per l), CD19+, 6-22% (0.05-0.61 x 10(9) per l), CD4+, 34-54% (0.45-1.65 x 10(9) per l), CD8+, 20-42% (0.29-1.17 x 10(9) per l), HLADR+, 4-23% (0.02-0.62 x 10(9) per l), CD56+, 4-22% (0.06-0.58 x 10(9) per l), CD45RO+, 16-53% (0.26-1.42 x 10(9) per l) and CD45RA+, 35-72% (0.34-2.05 x 10(9) per l). The mean CD4/CD8 ratio was 1.50+/-0.35. CD3+ cells were positively correlated to both CD4+ and CD8+ cells (P<0.001), and CD4+ cells showed a significant positive correlation with CD8+ cells (P<0.001).